Blastocystis species growth inhibition in vitro by plant extracts

Background: The protist Blastocystis species (sp.) inhabits the gastrointestinal tracts of humans and animals. In recent decades, alternative natural products derived from plants have demonstrated potential as effective treatments for Blastocystis infection. The anti-Blastocystis activity of three herbal ethanolic extracts— Odontites linkii subsp. cyprius, Ptilostemon chamaepeuce subsp. cyprius and Quercus alnifolia—were investigated in this study. Methods: Three distinct isolates of Blastocystis sp. maintained in vitro were molecularly subtyped. Cytotoxicity analysis was performed on individual Blastocystis sp. isolates using 250, 500, 1000, and 2000 μg/mL herbal ethanolic extracts for 24 and 48 hours. Quantitative, morphological, and size alterations of Blastocystis cells assessed the cytotoxicity of herbal anti-Blastocystis effect. Results: Following subtyping analysis, one strain of Blastocystis had ST3 and ST1 mixed subtypes, and two strains had ST1 subtypes. Starting after 24 h of incubation, P. cham. subsp. cyprius (1000 μg/mL) exhibited the most pronounced and consistent anti-Blastocystis cytotoxicity against all three strains, comparable to metronidazole. The Ptilostemon chamaepeuce subsp. cyprius anti-Blastocystis cytotoxicity was evident in parasite quantitative distress, morphological alterations, and significant reductions in cell size. Odontites linkii subsp. cyprius cytotoxicity varied among the three Blastocystis strains. The three Blastocystis strains were resistant to Quercus alnifolia. Conclusion: P. cham. subsp. cyprius was a potent and promising new herbal extract against Blastocystis sp. in vitro assays.